CN114712370A - Application of tomatidine in preparation of drug for resisting Israel acute paralysis virus - Google Patents
Application of tomatidine in preparation of drug for resisting Israel acute paralysis virus Download PDFInfo
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- CN114712370A CN114712370A CN202210491469.4A CN202210491469A CN114712370A CN 114712370 A CN114712370 A CN 114712370A CN 202210491469 A CN202210491469 A CN 202210491469A CN 114712370 A CN114712370 A CN 114712370A
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- tomatidine
- acute paralysis
- paralysis virus
- israel acute
- israel
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- XYNPYHXGMWJBLV-VXPJTDKGSA-N Tomatidine Chemical compound O([C@@H]1[C@@H]([C@]2(CC[C@@H]3[C@@]4(C)CC[C@H](O)C[C@@H]4CC[C@H]3[C@@H]2C1)C)[C@@H]1C)[C@@]11CC[C@H](C)CN1 XYNPYHXGMWJBLV-VXPJTDKGSA-N 0.000 title claims abstract description 61
- PWRIIDWSQYQFQD-UHFFFAOYSA-N sisunine Natural products CC1CCC2(NC1)OC3CC4C5CCC6CC(CCC6(C)C5CCC4(C)C3C2C)OC7OC(CO)C(OC8OC(CO)C(O)C(OC9OC(CO)C(O)C(O)C9OC%10OC(CO)C(O)C(O)C%10O)C8O)C(O)C7O PWRIIDWSQYQFQD-UHFFFAOYSA-N 0.000 title claims abstract description 61
- XYNPYHXGMWJBLV-OFMODGJOSA-N tomatidine Natural products O[C@@H]1C[C@H]2[C@@](C)([C@@H]3[C@H]([C@H]4[C@@](C)([C@H]5[C@@H](C)[C@]6(O[C@H]5C4)NC[C@@H](C)CC6)CC3)CC2)CC1 XYNPYHXGMWJBLV-OFMODGJOSA-N 0.000 title claims abstract description 61
- 241001163131 Israeli acute paralysis virus Species 0.000 title claims abstract description 46
- 239000003814 drug Substances 0.000 title claims abstract description 21
- 238000002360 preparation method Methods 0.000 title claims abstract description 13
- 229940079593 drug Drugs 0.000 title abstract description 8
- 241000700605 Viruses Species 0.000 claims description 11
- 241000208292 Solanaceae Species 0.000 claims description 4
- 239000007864 aqueous solution Substances 0.000 claims description 3
- 230000009385 viral infection Effects 0.000 claims description 3
- 239000002552 dosage form Substances 0.000 claims description 2
- 230000029812 viral genome replication Effects 0.000 claims description 2
- 230000001154 acute effect Effects 0.000 claims 2
- 208000018737 Parkinson disease Diseases 0.000 claims 1
- 239000004579 marble Substances 0.000 claims 1
- 241000317943 Acute bee paralysis virus Species 0.000 abstract description 12
- 230000000694 effects Effects 0.000 abstract description 12
- 230000010076 replication Effects 0.000 abstract description 6
- 208000015181 infectious disease Diseases 0.000 abstract description 5
- 230000005764 inhibitory process Effects 0.000 abstract description 3
- 241000257303 Hymenoptera Species 0.000 description 7
- 239000000243 solution Substances 0.000 description 6
- 238000000034 method Methods 0.000 description 4
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 3
- 241000227653 Lycopersicon Species 0.000 description 3
- 238000002474 experimental method Methods 0.000 description 3
- 230000006872 improvement Effects 0.000 description 3
- 238000003753 real-time PCR Methods 0.000 description 3
- 238000012360 testing method Methods 0.000 description 3
- WQZGKKKJIJFFOK-SVZMEOIVSA-N (+)-Galactose Chemical compound OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@H]1O WQZGKKKJIJFFOK-SVZMEOIVSA-N 0.000 description 2
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- 241000238631 Hexapoda Species 0.000 description 2
- 235000007688 Lycopersicon esculentum Nutrition 0.000 description 2
- 239000003513 alkali Substances 0.000 description 2
- 239000003443 antiviral agent Substances 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 239000010413 mother solution Substances 0.000 description 2
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- 230000010152 pollination Effects 0.000 description 2
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- 230000004083 survival effect Effects 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- GUAHPAJOXVYFON-ZETCQYMHSA-N (8S)-8-amino-7-oxononanoic acid zwitterion Chemical compound C[C@H](N)C(=O)CCCCCC(O)=O GUAHPAJOXVYFON-ZETCQYMHSA-N 0.000 description 1
- 241000238876 Acari Species 0.000 description 1
- 241000894006 Bacteria Species 0.000 description 1
- SRBFZHDQGSBBOR-IOVATXLUSA-N D-xylopyranose Chemical compound O[C@@H]1COC(O)[C@H](O)[C@H]1O SRBFZHDQGSBBOR-IOVATXLUSA-N 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- 241000233866 Fungi Species 0.000 description 1
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
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- 208000036142 Viral infection Diseases 0.000 description 1
- 229930013930 alkaloid Natural products 0.000 description 1
- 230000000507 anthelmentic effect Effects 0.000 description 1
- 230000000844 anti-bacterial effect Effects 0.000 description 1
- 230000000840 anti-viral effect Effects 0.000 description 1
- 244000052616 bacterial pathogen Species 0.000 description 1
- SRBFZHDQGSBBOR-UHFFFAOYSA-N beta-D-Pyranose-Lyxose Natural products OC1COC(O)C(O)C1O SRBFZHDQGSBBOR-UHFFFAOYSA-N 0.000 description 1
- 230000004071 biological effect Effects 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000008034 disappearance Effects 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 238000001917 fluorescence detection Methods 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 230000002458 infectious effect Effects 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 230000015788 innate immune response Effects 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 230000001717 pathogenic effect Effects 0.000 description 1
- 238000003752 polymerase chain reaction Methods 0.000 description 1
- 239000013641 positive control Substances 0.000 description 1
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- 108090000623 proteins and genes Proteins 0.000 description 1
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- 238000013215 result calculation Methods 0.000 description 1
- 238000010839 reverse transcription Methods 0.000 description 1
- 229930002534 steroid glycoside Natural products 0.000 description 1
- -1 steroidal glycoside alkaloid Chemical class 0.000 description 1
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- 238000010998 test method Methods 0.000 description 1
- 238000013518 transcription Methods 0.000 description 1
- 230000035897 transcription Effects 0.000 description 1
- 238000011144 upstream manufacturing Methods 0.000 description 1
Images
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/56—Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids
- A61K31/58—Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids containing heterocyclic rings, e.g. danazol, stanozolol, pancuronium or digitogenin
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/12—Antivirals
- A61P31/14—Antivirals for RNA viruses
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Abstract
The invention provides an application of tomatidine in preparation of an anti-israel acute paralysis virus medicine, and relates to the technical field of biological medicines. The invention firstly provides that the tomatidine has a definite effect of resisting the Israel acute paralysis virus, can obviously inhibit bee death caused by infection of the Israel acute paralysis virus, can obviously inhibit the replication of the Israel acute paralysis virus, and has an obvious inhibition effect on the replication of the RNA of the Israel acute paralysis virus at the concentration of 10uM of the tomatidine, which shows that the tomatidine has a certain effect of resisting the Israel acute paralysis virus and has a good prospect when being applied to the preparation of the anti-Israel acute paralysis virus medicine.
Description
Technical Field
The invention belongs to the technical field of biological medicines, and particularly relates to application of tomatidine in preparation of an anti-israel acute paralysis virus medicine.
Background
As social insects, bees make an important contribution to plant pollination, and about 35% of crops need to rely on the pollination to a certain extent in the global range. However, events of bee colony reduction occur to varying degrees worldwide. Especially in the winter to spring of the year, a wide range of sudden bee disappearance occurs in the united states, and the same phenomenon occurs in parts of bee farms in europe, which scientists define as Colony Collapse Disorder (CCD). It is now generally accepted that a synergistic interaction between factors (bee mites, chlordime, viruses, etc.) results in a large loss of bee colonies, the major virus being the Israel Acute Paralysis Virus (IAPV).
Since Israel acute paralysis virus is not only highly pathogenic, but also highly variable, bees can rely only on innate immunity, and antiviral drugs play an important role in protecting bee colonies and controlling viral infection. With the development of modern molecular biology technology, research on antiviral drugs has been greatly advanced, however, up to now, the anti-israel acute paralysis virus drug is still a candidate drug to be developed, which has disadvantages, is difficult to obtain ideal antiviral effect, and the search and discovery of a new anti-israel acute paralysis virus drug are repeated and far away.
Tomatidine (Tomatidine) is a steroidal glycoside alkaloid containing D-xylose, D-galactose and two molecules of glucose. The chemical structural formula is as follows:
according to previous researches, the tomatidine can protect tomato plants from being invaded by bacteria, fungi, viruses and certain insects in the growth process, and has obvious antibacterial and anthelmintic effects. According to literature reports, the tomatidine can inhibit some pathogenic bacteria harmful to human and animals, and has remarkable biological activity. However, the anti-Israel acute paralysis virus effect of the tomatidine and the application of the tomatidine in the preparation of the anti-Israel acute paralysis virus medicine are not reported at home and abroad, and related documents are not retrieved.
Disclosure of Invention
The invention provides an application of tomatidine in preparation of an anti-israel acute paralysis virus medicine, overcomes the defects of the prior art, and can effectively solve the problems that the anti-israel acute paralysis virus effect of the tomatidine and the application of the tomatidine in preparation of the anti-israel acute paralysis virus medicine are not reported at home and abroad.
In order to achieve the aim, the invention provides an application of tomatidine in resisting Israel acute paralysis virus.
The following is further optimization or/and improvement of the technical scheme of the invention:
the tomatidine can obviously inhibit the RNA expression of the Israel acute paralysis virus.
The tomatidine inhibits israel acute paralysis virus replication.
The tomatidine inhibits bee death caused by Israel acute paralysis virus infection.
The concentration of the tomatidine is 10 uM.
In addition, the invention also provides an application of the tomatidine in preparing the anti-israel acute paralysis virus medicine.
The following is further optimization or/and improvement of the technical scheme of the invention:
the tomatidine is extracted from Solanaceae plants;
the Solanaceae plant comprises fructus Lycopersici Esculenti.
The dosage form of the tomatidine is aqueous solution.
The concentration of the tomatidine is 10 uM.
The invention has the following advantages:
the invention firstly provides that the tomatidine has a definite effect of resisting the Israel acute paralysis virus, can obviously inhibit bee death caused by infection of the Israel acute paralysis virus, and meanwhile, the tomatidine can obviously inhibit replication of the Israel acute paralysis virus and reduce the expression level of RNA of the Israel acute paralysis virus, so that the tomatidine has the effect of resisting the Israel acute paralysis virus and has good prospects when being applied to preparation of the medicament for resisting the Israel acute paralysis virus.
Drawings
FIG. 1 is a graph of survival rate of bees infected with Israel acute paralysis virus after treatment with tomatidine.
FIG. 2 shows the Real-time PCR detection of the expression of the tomatidine on the Israel acute paralysis virus RNA.
Detailed Description
The following examples further illustrate the invention but are not intended to limit the invention thereto.
The tomatidine in the invention can be sold in the market, the tomatidine solution can be prepared according to the existing method, the tomatidine is dissolved by DMSO, then the tomatidine solution is prepared into mother solution with the concentration of 4.16mg/ml, and the mother solution is added into 999ml of water and evenly mixed. The concentration of tomatidine was 4.16mg by weight per liter of solution, i.e. 10 uM. The clinical dosage is 100-.
Example 1
Effect of tomatidine on bee survival rate of infection with israel acute paralysis virus:
experimental materials: israel Acute Paralysis Virus (IAPV), which is obtained by in vitro transcription after IAPV infectious clone is constructed in the laboratory and is preserved at the temperature of minus 80 ℃. In the experiment, the aqueous solution is prepared into solution with required concentration for administration, and the administration concentration or dosage is calculated according to the raw material of the tomatidine.
The experimental principle is as follows: italian bees are taken as virus hosts, and the death condition of the bees and the RNA replication condition of Israel acute paralysis virus are determined.
Effect of tomatidine samples on bee death infected with israel acute paralysis virus: after the bees are inoculated with the viruses, the bees are placed at 50% humidity and cultured at 30 ℃. Spraying tomato alkali solution after 24 hr for 2 times a day for 3 days. Setting virus control and negative control at the same time, culturing at 30 deg.C. The culture was continued for 9 days. The results of the experiment are shown in FIG. 1.
And (4) conclusion: as can be seen from FIG. 1, under the test conditions, the tomatidine has better inhibitory activity to the Israel acute paralysis virus, the result of the positive control drug is consistent with the previous result in the laboratory, the test system is established, and the test result is credible.
Example 2
The inhibition effect of the tomatidine on the israel acute paralysis virus RNA:
the test method comprises the following steps:
after the bee is inoculated with virus, the bee is placed in 50% humidity and cultured at 30 ℃. Spraying tomato alkali solution after 24 hr for 2 times a day for 3 days. Virus control and negative control were set simultaneously, and cultured at 30 ℃. The culture was continued for 9 days.
Real–time PCR：
Bee samples were ground with liquid nitrogen and total RNA was extracted by trizol method. RNA was quantified at 1ug and reverse transcribed. After the reverse transcription is finished, the sample is diluted by 10 times, and 1ul is taken for subsequent experiments.
Designing and synthesizing specific primers:
an upstream primer: 5'-CCAGCCGTGAAACATGTTCTTACC-3' the flow of the air in the air conditioner,
a downstream primer: 5'-ACATAGTTGCACGCCAATACGAGAAC-3' the flow of the air in the air conditioner,
fluorescence quantification using KAPA SYBR FAST qPCR kit, reaction system: 7.5. mu.L Mix, 0.3. mu.L Forward Primer (10. mu.M), 0.3. mu.L reverse Primer (10. mu.M), 1. mu.L Template (100pg to 1. mu.g of genomic RNA), and 5.9. mu.L Water. Reaction conditions are as follows: 95 ℃ for 3min, 40cycles of: 95 ℃ for 3s and 60 ℃ for 30 s.
Q-PCR absolute quantitative result calculation method: the Bori fluorescence detection system is adopted to carry out absolute quantitative analysis on the target gene, and Real-time PCR (polymerase chain reaction) is used for detecting the replication of the tomatidine to the Israel acute paralysis virus, which is shown in figure 2.
The experimental results are as follows: as can be seen from fig. 2, the IAPV copy number at the dose of tomatidine 10uM was significantly reduced compared to the virus control group, and the results indicate that tomatidine could significantly inhibit israel acute paralysis virus RNA replication.
In conclusion, the invention firstly provides that the tomatidine has a definite effect of resisting the Israel acute paralysis virus, can obviously inhibit bee death caused by infection of the Israel acute paralysis virus, has an obvious inhibition effect on the expression level of RNA of the Israel acute paralysis virus at the concentration of 10uM of the tomatidine, and has a good prospect when being applied to the preparation of the anti-Israel acute paralysis virus medicine.
The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention, and various modifications and changes may be made by those skilled in the art. Any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the protection scope of the present invention.
Claims (9)
1. An application of tomatidine in resisting Israel acute paralysis virus is provided.
2. The use of tomatidine according to claim 1 against the israel acute paralysis virus, wherein: the tomatidine can obviously inhibit the RNA expression of the Israel acute paralysis virus.
3. The use of tomatidine according to claim 1 against the israel acute paralysis virus, wherein: the tomatidine inhibits israel acute paralysis virus replication.
4. The use of tomatidine according to claim 1 against the israel acute paralysis virus, wherein: the tomatidine inhibits bee death caused by Israel acute paralysis virus infection.
5. The use of tomatidine according to claim 1 against the israel acute paralysis virus, wherein: the concentration of the tomatidine is 10 uM.
6. An application of tomatidine in preparing medicine for treating acute paralysis agitans virus is disclosed.
7. The use of tomatidine as in claim 6 for the preparation of a medicament against Israel acute Marble's Virus, wherein: the tomatidine is extracted from Solanaceae plants;
the Solanaceae plant comprises fructus Lycopersici Esculenti.
8. The use of tomatidine as in claim 6 or 7 in the preparation of a medicament against israel acute paralysis virus, wherein: the dosage form of the tomatidine is aqueous solution.
9. The use of tomatidine as in claim 8 for the preparation of a medicament against israel acute paralysis virus, wherein: the concentration of the tomatidine is 10 uM.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210491469.4A CN114712370A (en) | 2022-05-07 | 2022-05-07 | Application of tomatidine in preparation of drug for resisting Israel acute paralysis virus |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210491469.4A CN114712370A (en) | 2022-05-07 | 2022-05-07 | Application of tomatidine in preparation of drug for resisting Israel acute paralysis virus |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN114712370A true CN114712370A (en) | 2022-07-08 |
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202210491469.4A Pending CN114712370A (en) | 2022-05-07 | 2022-05-07 | Application of tomatidine in preparation of drug for resisting Israel acute paralysis virus |
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| CN (1) | CN114712370A (en) |
Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1344169A (en) * | 1999-02-26 | 2002-04-10 | 坎塔布连制药工业股份有限公司 | Glycoconjugates obtained from i(candide Utilis) cells and i(Ricinus)i(communis) seeds |
| US20120157512A1 (en) * | 2009-08-21 | 2012-06-21 | Monsanto Technology Llc | Preventing and Curing Beneficial Insect Diseases Via Plant Transcribed Molecules |
| KR20130112128A (en) * | 2012-04-03 | 2013-10-14 | 대한민국(농촌진흥청장) | Composition for immune enhancement of honey bee |
| CN105980399A (en) * | 2013-12-10 | 2016-09-28 | 比欧洛吉科斯公司 | Compositions and methods for virus control in varroa mite and bees |
| WO2019004825A1 (en) * | 2017-06-28 | 2019-01-03 | Rijksuniversiteit Groningen | Tomatidine and analogs thereof for use as antiviral agent |
| CN110638871A (en) * | 2019-10-12 | 2020-01-03 | 中国农业科学院蜜蜂研究所 | Anti-bee paralysis virus extract and preparation method thereof |
| CN111920821A (en) * | 2020-03-12 | 2020-11-13 | 南京农业大学 | Application of tomatidine in preparation of antiviral drugs |
-
2022
- 2022-05-07 CN CN202210491469.4A patent/CN114712370A/en active Pending
Patent Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1344169A (en) * | 1999-02-26 | 2002-04-10 | 坎塔布连制药工业股份有限公司 | Glycoconjugates obtained from i(candide Utilis) cells and i(Ricinus)i(communis) seeds |
| US20120157512A1 (en) * | 2009-08-21 | 2012-06-21 | Monsanto Technology Llc | Preventing and Curing Beneficial Insect Diseases Via Plant Transcribed Molecules |
| KR20130112128A (en) * | 2012-04-03 | 2013-10-14 | 대한민국(농촌진흥청장) | Composition for immune enhancement of honey bee |
| CN105980399A (en) * | 2013-12-10 | 2016-09-28 | 比欧洛吉科斯公司 | Compositions and methods for virus control in varroa mite and bees |
| WO2019004825A1 (en) * | 2017-06-28 | 2019-01-03 | Rijksuniversiteit Groningen | Tomatidine and analogs thereof for use as antiviral agent |
| CN110638871A (en) * | 2019-10-12 | 2020-01-03 | 中国农业科学院蜜蜂研究所 | Anti-bee paralysis virus extract and preparation method thereof |
| CN111920821A (en) * | 2020-03-12 | 2020-11-13 | 南京农业大学 | Application of tomatidine in preparation of antiviral drugs |
Non-Patent Citations (2)
| Title |
|---|
| WANG, PENGCHENG，等: "Tomatidine inhibits porcine epidemic diarrhea virus replication by targeting 3CL protease", 《VETERINARY RESEARCH》, vol. 51, no. 1, pages 1 - 18 * |
| 张雪琦，等: "中华蜜蜂囊状幼虫病发病及防治研究进展", 《应用昆虫学报》, vol. 51, no. 04, pages 11 - 12 * |
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